In our country, chemical methods are frequently used in the fight against agricultural diseases since they take effect in a short period of time and yield quick results. Pesticide use in Turkey has increased by 45% in the last 20 years. Pesticides are widely used especially in Mediterranean and Aegean regions of Turkey. Unconscious and uncontrolled use of pesticides has adverse effects on the environment and human health since they lead to development of persistence and residues in harmful microorganisms. Decrease or elimination of synthetic pesticide use in agriculture is desired. The most promising means to accomplish this goal is using new techniques based on biological control agents (BCAs) for disease control and, consequently, minimizing chemicals' detrimental effects on the environment.
Other methods, especially biological control, have been gaining prominence due to the fact that chemical control, one of the plant protection methods in agricultural production, has adverse effects on the environment and humans and that pathogens develop persistence against these. Biological control agents, including fungi, find a wide field of use along with bacteria (mainly Bacillus thuringiensis) since they have a wide spectrum in disease control, and owing, to their productive efficiency.
Fungi types such as Trichoderma spp., Gliocladium spp., Aspergillus spp., Alternaria spp., Fusarium spp., Chaetomium spp., Ampelomyces spp., Coniothyrium spp., Sporidosumun spp. and Teretosperma spp. provide significant antagonist effect. Moreover, Trichoderma species has a 50% share in fungal biological control agent market rather as a soil/growth enhancer which has led to more research being conducted on Trichoderma species.
Depending on the strain, Trichoderma use in agriculture provides numerous advantages. These advantages are: (i) biological control agent building colonies in rhizosphere allows it to settle quickly in constant microbial communities; (ii) pathogenic and competitive/harmful microflora is controlled by using various mechanisms; (iii) plant health is enhanced and (iv) root development is stimulated. As Trichoderma based biological control agents are compared with their counterparts (virus, bacteria, nematodes and protozoa), they come to the fore due to reasons such as enhancing plant growth and exhibiting high level of activity in remediation of soil.
The ultimate goal for production of any biological control agent is to implement the most economical mass production. Almost all bio-control agent products that are based on Trichoderma species contain spores as the active content.
This is associated with the physiological conditions of three microbial propagules (mycelia, conidia and chlamydospores) of Trichoderma species.
These three propagules have different physiological characteristics with regard to production, stability and bio-control agent activity. Thus, it is essential to choose propagules with the most suitable structures in order for Trichoderma species function as bio-control agents in an efficient manner.
Although exhibit perfect bio-control agent activity, mycelia, cannot survive downstream process steps such as drying and thus are not practical. Chlamydospores, on the other hand, require a 2-3 weeks period for cultivation and, although they are more persistent than mycelia, cannot survive drying process just like mycelia. As mentioned above, conidia are active bio-control agents, affected less by various environmental, conditions and may be produced in larger amounts in a much shorter period of 3-4 days. However, mycelia are inevitably present in the production environment along with conidia. Moreover, simultaneous production of mycelia ensures that there are various basic metabolites (e.g. antibiotics) for bio-control agent activity. For this reason, production of micropropagules containing conidia as lain propagules in addition to mycelia is the best strategy of producing Trichoderma species.
While commercial Trichoderma species preparations for biological control generally consist of bulk conidia, a good bio-control activity depends on fungus remaining vegetative, that is, being antagonistically active. For this reason, effective and efficient use of Trichoderma based biological control agents involves establishing a balance between formation of conidia at a very low cost during, production and drastic vegetative growth during use.
Commercial success of bio-control agents based on Trichoderma species also requires economically suitable biomass production processes (35-40% of production costs depends on raw material)
Despite its advantages, biomass production of Trichoderma based biological control agents is not very common, since high cost materials are used in production such as Mendelian conditions, molasses and corn steep liquor. The cost of such raw materials used in commercial production of biological control agents is the primary limiting factor. In order to overcome cost limitation, a number of researchers have successfully utilized dry corn fiber mass, sewage sludge compost and cranberry pulp. Despite use of alternative resources, production cost remains high due to the fact that these raw materials should be supported by other nutrients.
Growth and production are influenced by numerous parameters such as culture medium, inoculation, pH, temperature, oxygen and mass transfer. It is indicated that since filamentous fungi such as Trichoderma have various morphological structures, optimization and scaling of fermentation processes are difficult. Commercial production of conidia depends generally on manipulation of nutrients and substrata that support conidiation. Numerous studies have been conducted on optimal growth conditions for ensuring in vitro, conidia formation in many Trichoderma species. These studies have indicated that, in addition to medium pH, carbon and nitrogen conditions and C:N ratio are the primary environmental factors affecting conidia formation in Trichoderma. Although aerial spores (conidia) may be produced in solid media, in further phases of the process, large scale production of conidia becomes difficult due to the fact that optimal temperature and ventilation values cannot be maintained. It is known that liquid fermentation is used for chlamydospores production. However, not every fungus has the capability of producing spores in deep culture. For this reason, production conditions should be optimized specifically for the chosen species in order to obtain maximum propagule yield.
For mass production of a biological agent by means of solid culture fermentation, a very large amount of spore biomass is required. Substrates such as grains, unsieved coarse flour, brans, hay, plant residues and organic fertilizers are used for mass production of Trichoderma harzianum. Pulp residue (even used pulp residue) is a suitable support substrate for solid culture fermentation. Wine pulp contains various nutrients for microbial growth such as carbon resource, nitrogen resource and trace elements. Such wine wastes are substantially inexpensive substrates and are suitable for solid culture fermentation. Moreover, other agricultural-industrial wastes such as solid paper wastes and cellulosic plant residues from paper factories are released to the environment every year, increase pollution and lead to waste disposal problems. For this reason, in line with the need for large scale low cost production of environmentally friendly bio-pesticides, studies for regionally available more inexpensive substrates are being conducted for mass production of T. harzianum. 
Although there are no patents in the field related to Trichoderma citrinoviride strain, there are patents that covers production of Trichoderma harzianum conidia which is of the same genus and whose patent subject matter relates to production, namely U.S. Pat. No. 5,330,912 A 19940719, U.S. Pat. No. 5,422,107 A 19950606 and WO 2007/094014 A1 20070823.